ArticleScientific reports2018
Engineered membrane protein antigens successfully induce antibodies against extracellular regions of claudin-5.
Article in Scientific reports, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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Who cites it
20 citing papers in PubMed, 37 citations in OpenAlex.
- Multi-dimensional orchestration of binders for improved CAR-T immunotherapy.Cancer letters · 2026Review
- Structure-forward targeting of claudins with synthetic binders.FEBS letters · 2026Review
- Design of a water-soluble CD20 antigen with computational epitope scaffolding.Protein science : a publication of the Protein Society · 2025Article
- A claudin5-binding peptide enhances the permeability of the blood-brain barrier in vitro.Science advances · 2025Article
- Advancements in the conservation of the conformational epitope of membrane protein immunogens.Frontiers in immunology · 2025Review
- Association of Plasma Claudin-5 with Age and Alzheimer Disease.International journal of molecular sciences · 2024Article
- Production of Immunizing Antigen Proteoliposome Using Cell-Free Protein Synthesis System.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Storming the gate: New approaches for targeting the dynamic tight junction for improved drug delivery.Advanced drug delivery reviews · 2023Review
- Bortezomib Increased Vascular Permeability by Decreasing Cell-Cell Junction Molecules in Human Pulmonary Microvascular Endothelial Cells.International journal of molecular sciences · 2023Article
- Article
- Evaluation of Cell-Free Synthesized Human Channel Proteins for In Vitro Channel Research.Membranes · 2022Article
- Improving antibody drug development using bionanotechnology.Current opinion in biotechnology · 2022Review
- Modulation of the Blood-Brain Barrier for Drug Delivery to Brain.Pharmaceutics · 2021Review
- Cell-free expression of the outer membrane protein OprF ofLife science alliance · 2021Article
- Therapeutic Nanobodies Targeting Cell Plasma Membrane Transport Proteins: A High-Risk/High-Gain Endeavor.Biomolecules · 2021Review
- Tight Junction Modulating Bioprobes for Drug Delivery System to the Brain: A Review.Pharmaceutics · 2020Review
- Non-full-length Water-Soluble CXCR4iScience · 2020Article
- Cell-Free Protein Synthesis: A Promising Option for Future Drug Development.BioDrugs : clinical immunotherapeutics, biopharmaceuticals and gene therapy · 2020Review
- Potential for Tight Junction Protein-Directed Drug Development Using Claudin Binders and Angubindin-1.International journal of molecular sciences · 2019Review
- Autoimmune receptor encephalitis in mice induced by active immunization with conformationally stabilized holoreceptors.Science translational medicine · 2019Article
Corrections and comments
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Authors and funding
10 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The production of antibodies against the extracellular regions (ECR) of multispanning membrane proteins is notoriously difficult because of the low productivity and immunogenicity of membrane proteins due to their complex structure and highly conserved sequences among species. Here, we introduce a new method to generate ECR-binding antibodies utilizing engineered liposomal immunogen prepared using a wheat cell-free protein synthesis system. We used claudin-5 (CLDN-5) as the target antigen, which is a notoriously difficult to produce and poorly immunogenic membrane protein with two highly conserved extracellular loops. We drastically improved the productivity of CLDN-5 in the cell-free system after suppressing and normalizing mRNA GC content. To overcome its low immunogenicity, two engineered antigens were designed and synthesized as proteoliposomes: a human/mouse chimeric CLDN-5, and a CLDN-5-based artificial membrane protein consisting of symmetrically arranged ECRs. Intraperitoneal immunization of both engineered CLDN-5 ECR antigens induced ECR-binding antibodies in mice with a high success rate. We isolated five monoclonal antibodies that specifically recognized CLDN-5 ECR. Antibody clone 2B12 showed high affinity (<10 nM) and inhibited CLDN-5-containing tight junctions. These results demonstrate the effectiveness of the methods for monoclonal antibody development targeting difficult-to-produce membrane proteins such as CLDNs.
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